Novel evidence for an ecto-phospholipid methyltransferase in isolated rat hepatocytes.
Phospholipids of isolated rat hepatocytes were labelled by preincubation with either 2 microM -methyl-14C-S-adenosylmethionine (AdoMet) or 2 microM [methyl-14C]methionine. Subsequent addition of phospholipase C to the suspension removed 95% of the radioactivity from phospholipids methylated by [methyl-14C]AdoMet within a few minutes, but was without effect on phospholipids methylated by [methyl-14C]methionine radioactivity from the latter could, nevertheless, be removed by phospholipase C after permeabilization of the cells with digitonin. The results clearly show that the methyl group of exogenous AdoMet, contrary to that of methionine, is transferred on to phospholipids located on the external face of the plasma membrane. Accordingly, pretreatment of isolated hepatocytes with trypsin prevented the methylation of phospholipids from exogenous AdoMet by 60-80%, whereas it was almost without effect when exogenous methionine was the methyl donor. Our data corroborate previous work [Bontemps and Van den Berghe (1997) Biochem. J. 327, 383-389], which indicated that AdoMet methylates hepatocyte phospholipids without penetrating the cells.
Bontemps
F
F
Laboratory of Physiological Chemistry, International Institute of Cellular and Molecular Pathology, and University of Louvain Medical School, Avenue Hippocrate 75, B-1200 Brussels, Belgium.
Van Den Berghe
G
G
D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D005110:Extracellular Space / Q000201:enzymology; D008099:Liver / Q000201:enzymology*; D008297:Male; D008563:Membrane Lipids / Q000378:metabolism*; D008745:Methylation; D008780:Methyltransferases / Q000378:metabolism*; D050918:Phosphatidylethanolamine N-Methyltransferase; D051381:Rats; D017208:Rats, Wistar; D012436:S-Adenosylmethionine / Q000378:metabolism; D010738:Type C Phospholipases / Q000378:metabolism
D008563:Membrane Lipids; D012436:S-Adenosylmethionine; D008780:Methyltransferases; D050918:Phosphatidylethanolamine N-Methyltransferase; D010738:Type C Phospholipases